| RRC ID |
38134
|
| Author |
Yamada M, Hori A, Sugaya S, Yajima Y, Utoh R, Yamato M, Seki M.
|
| Title |
Cell-sized condensed collagen microparticles for preparing microengineered composite spheroids of primary hepatocytes.
|
| Journal |
Lab Chip
|
| Abstract |
The reconstitution of extracellular matrix (ECM) components in three-dimensional (3D) cell culture environments with microscale precision is a challenging issue. ECM microparticles would potentially be useful as solid particulate scaffolds that can be incorporated into 3D cellular constructs, but technologies for transforming ECM proteins into cell-sized stable particles are currently lacking. Here, we describe new processes to produce highly condensed collagen microparticles by means of droplet microfluidics or membrane emulsification. Droplets of an aqueous solution of type I collagen were formed in a continuous phase of polar organic solvent followed by rapid dissolution of water molecules into the continuous phase because the droplets were in a non-equilibrium state. We obtained highly unique, disc-shaped condensed collagen microparticles with a final collagen concentration above 10% and examined factors affecting particle size and morphology. After testing the cell-adhesion properties on the collagen microparticles, composite multicellular spheroids comprising the particles and primary rat hepatocytes were formed using microfabricated hydrogel chambers. We found that the ratio of the cells and particles is critical in terms of improvement of hepatic functions in the composite spheroids. The presented methodology for incorporating particulate-form ECM components in multicellular spheroids would be advantageous because of the biochemical similarity with the microenvironments in vivo.
|
| Volume |
15(19)
|
| Pages |
3941-51
|
| Published |
2015-10-7
|
| DOI |
10.1039/c5lc00785b
|
| PMID |
26308935
|
| MeSH |
Animals
Cell Adhesion
Cells, Cultured
Collagen Type I / chemistry*
Cytochrome P-450 CYP3A / genetics
Cytochrome P-450 CYP3A / metabolism
Hep G2 Cells
Hepatocytes / cytology*
Hepatocytes / metabolism
Humans
Hydrogel, Polyethylene Glycol Dimethacrylate / chemistry
Mice
Microfluidic Analytical Techniques / instrumentation
Microfluidic Analytical Techniques / methods
Microscopy, Fluorescence
Microspheres*
NIH 3T3 Cells
Ornithine Carbamoyltransferase / genetics
Ornithine Carbamoyltransferase / metabolism
Rats
Real-Time Polymerase Chain Reaction
Spheroids, Cellular / cytology*
Spheroids, Cellular / metabolism
|
| IF |
6.774
|
| Times Cited |
37
|
|
WOS Category
|
BIOCHEMICAL RESEARCH METHODS
CHEMISTRY, MULTIDISCIPLINARY
NANOSCIENCE & NANOTECHNOLOGY
|
| Resource |
| Human and Animal Cells |
NIH3T3
Hep G2 |
